Method for the devitalisation of natural organs and/or for the preparation of extracellular matrices for tissue engineering

Abstract

74 . Culture medium according to any one of claims 48 to 73 , characterized in that, that autologous growth factors are gained through chemical and/or biochemical destruction of body-own tissues. 75 . Culture medium according to any one of claims 48 to 74 , characterized in that, that autologous growth factors are gained through apoptosis of body-own tissues. 76 . Culture medium according to claim 74, characterized by the fact that tissue destruction is done by ultrasound. Summary The invention provides new procedures for the devitalization and preserving of human and animal organs and tissues, preferably however natural hollow organs and all of their components, in particular from blood vessels and cardiac valves. Furthermore the invention provides procedures for the production of matrices for construction of organs and tissues in part or in toto. In addition the invention concerns organs and tissues, in particular natural and artificial hollow organs which can be achieved according to invention-appropriate procedures. Furthermore the invention concerns the clinical use of these organs and tissues an the application in the human and veterinarian medicine, preferably in the cardiac and vascular surgery. Furthermore the invention concerns new culture media.

1 . Procedure or process for devitalization and preservation of organs and/or tissues, comprising the following steps: 
 a. sterile harvest and storage of the organ or the tissue in a liquid selected from the group consisting of: sterile water, a crystalloid liquid, a colloidal liquid, a lipid-containing liquid or a combination of the mentioned liquids, until the devitalization is achieved,    b. the outwash of cell-fragments, cellular decomposition products (debris) as well as soluble substances under pressure with a liquid, selected from the group consisting of: sterile water, a crystalloid liquid, a colloidal liquid, a lipid-containing liquid and a combination of the mentioned liquids.    
     
     
         2 . The procedure or process of  claim 1  wherein the sterile harvest of the organ and/or tissue is performed from dead people (multi organ donors).  
     
     
         3 . The procedure or process of  claim 1  wherein the storage occurs for at least 2 weeks, preferably for 6 weeks, in particular preferred however for 6 months in the dark.  
     
     
         4 . The procedure or process of  claim 1  wherein the storage occurs under sterile conditions.  
     
     
         5 . The procedure or process of  claim 1  to  4 , wherein the storage liquid is a crystalloid liquid.  
     
     
         6 . The procedure or process of  claim 5  wherein the crystalloid liquid is Medium 199 (Seromed).  
     
     
         7 . The procedure or process of  claim 5  wherein the crystalloid liquid is cardioplegic solution (Bretschneider solution).  
     
     
         8 . The procedure or process of  claim 5  wherein the crystalloid liquid is supplemented with antibiotics.  
     
     
         9 . The procedure or process of  claim 1  to  7  wherein the procedure contains a multiple rinsing of the organs and/or tissues before the storage in the same liquid in which the storage occurs.  
     
     
         10 . The procedure or process of  claim 1  to  8  wherein the storage and the outwash of the organs and/or tissues occurres with the same liquid.  
     
     
         11 . The procedure or process of  claim 1  to  9  wherein the storage is done at a pH-value from 3 to 9.  
     
     
         12 . The procedure or process of  claim 10  wherein the storage is done at a pH-value from 6.9 to 7.8.  
     
     
         13 . The procedure or process of  claim 10  wherein the storage is done at a pH-value from 7.0 to 7.5.  
     
     
         14 . The procedure or process of  claim 1  to  12  wherein the storage is done at a temperature of 0 to 55° C.  
     
     
         15 . The procedure or process of  claim 13  wherein the storage is done at a temperature of 0 to 37° C.  
     
     
         16 . The procedure or process of  claim 13  wherein the storage is done at a temperature of 4° C.  
     
     
         17 . The procedure or process of  claim 1  to  15  wherein the storage is done under reduced oxygen pressure.  
     
     
         18 . The procedure or process of  claim 16  wherein the storage is done under anaerobe conditions.  
     
     
         19 . The procedure or process of  claim 1  to  15  wherein the storage is done with gases (in the fluid or gaseous stage).  
     
     
         20 . The procedure or process of  claim 18  wherein the gas is a rare gas.  
     
     
         21 . The procedure or process of  claim 1  to  20  wherein the washing out occurs pulsating.  
     
     
         22 . The procedure or process of  claim 21  wherein the washing out of cellular particles, cellular decomposition products, as well as soluble substances is done repeatedly.  
     
     
         23 . The procedure or process of  claim 22  wherein the washing out is done at least twice, preferably at intervals of 6 weeks.  
     
     
         24 . The procedure or process of  claim 1  wherein invention-appropriately treated organs and tissues are dried after devitalization and preservation.  
     
     
         25 . The procedure or process of  claim 1  wherein the organs are hollow organs.  
     
     
         26 . The procedure or process of  claim 1  wherein the washing out is performed by use of a cultivation apparatus comprising a culture vessel which is filled with a liquid ( 1 ), two adapters ( 3 ) that are connected with the two outflows of the culture vessel, a hose, that is connected with a pump and a further hose that ends at a stock or dump vessel ( 5 ) wherein the pressure gradient Δp is dependent on the riser tube ( 4 ) and pressure transducer ( 8 ).  
     
     
         27 . The procedure or process of  claim 1  wherein the washing out is done by use of a cultivation apparatus in accordance with  FIG. 1 .  
     
     
         28 . The procedure or process of  claim 1  wherein the organs and tissues are selected from the group consisting of: Blood vessels, blood vessel valves, lymphatic vessels, lymphatic vessel valves, ureters, bladders, spermatic ducts, bronchial tubes, livers, kidneys, hearts and cardiac valves.  
     
     
         29 . Organs and tissues produced by the procedure or process of  claim 1  to  27 .  
     
     
         30 . The procedures or processes for the production of matrices for the partial or denovo synthesis of organs and/or tissues comprising the following steps: 
 a. devitalization and preservation of organs and/or tissues according to the procedures or processes of one of the  claims 1  to  27 ,    b. cell-repopulation of the organs and tissues, preferably through re-endothelialization.    
     
     
         31 . The procedure or process of  claim 29  wherein the organs are hollow organs.  
     
     
         32 . The procedure or process of caim  30  wherein the hollow organs are selected from the group consisting of: allogenic vessels, to xenogenic vessels and ureter.  
     
     
         33 . Organs produced which are produced according to one of the  claims 29  to  31 .  
     
     
         34 . The procedure or process of  claim 29  wherein the cells used for re-population are autologous cells, for example endothelial cells.  
     
     
         35 . The procedure or process of  claim 29  wherein the re-endothelialization is done by the seeding of cells on the intraluminal surface of the hollow organs.  
     
     
         36 . The procedure or process of  claim 29  wherein a precoating of the hollow organs with adhesion molecules is performed before cell seeding.  
     
     
         37 . The procedure or process according to  claim 29 , characterized in that the hollow organ is precoated with autologous serum before seeding of the cells.  
     
     
         38 . The procedure or process according to  claim 30 , characterized in that the hollow organ is precoated with autologous growth and adhesion factor rich serum before seeding of the cells.  
     
     
         39 . The procedure or process according to  claim 30 , further comprising natural and artificial hollow organs selected from the group consisting from: blood vessels, blood vessel valves, lymphatic vessels, lymphatic vessel valves, ureters, bladders, spermatic ducts, bronchial tubes, hearts and cardiac valves.  
     
     
         40 . Use of the tissues or organs, available through the procedure or process according to one of the  claims 1  to  28  in the medicine and veterinarian medicine.  
     
     
         41 . Use of the vessels, available through the procedure or process according to one of the  claims 1  to  28  in the cardiac and vascular surgery.  
     
     
         42 . Use of the vessels, available through the procedure or process according to one of the  claims 1  to  28  as an aortocoronary bypass or as a vascular transplant in vascular reconstructions.  
     
     
         43 . Use of the vessels, available through the procedure or process according to one of the  claims 1  to  28  in peripheral arterial disease, aneurysms, redooperations in cardiac and vascular surgery and in the case of innate deformations of the vascular system.  
     
     
         44 . Tissues and organs, available through the procedure or process according to one of the  claims 1  to  28 , characterized in that the tissue and organ is in addition surrounded by a coat of a synthetic material.  
     
     
         45 . Tissues and organs, available through the procedure or process according to one of the  claims 1  to  29 , characterized in that the tissue and organ is in addition surrounded by a coat of a synthetic resorbable material.  
     
     
         46 . Tissues and organs, available through the procedure or process according to one of the  claims 1  to  28 , characterized in that the tissue and organ is in addition surrounded by a coat of a synthetic resorbable material, comprising polyglycon acide.  
     
     
         47 . Cultivation device (bioreactor) for the use in a procedure or process according to  claim 1 , comprising a culture vessel filled with a solution ( 1 ), two adapters ( 3 ) which are connected with the two outflows of the culture vessel, a hose, that is connected with a computer controlled peristaltic pump and a further hose that ends at an stock or dump vessel ( 5 ). The pressure gradient Δp is dependent on the riser tube ( 4 ) and pressure transducer ( 8 ).  
     
     
         48 . Culture medium for the increase of growth, remodeling processes and reduction of dedifferenziation of vascular cells in tissue culture, characterized in that autologous growth factors and/or adhesion molecules are added to a basal chemically defined medium or to a full medium.  
     
     
         49 . Culture medium according to  claim 48 , characterized in that the growth factors and/or adhesion molecules are added in autologous, not heat inactivated serum.  
     
     
         50 . Culture medium according to  claim 48 , comprising 5-30% of autologous serum.  
     
     
         51 . Culture medium according to  claim 48 , comprising 5-20% autologous serum.  
     
     
         52 . Culture medium according to  claim 48 , comprising 10-15% autologous serum.  
     
     
         53 . Culture medium according to any of the  claims 48  to  52 , characterized in that additionally recombinant growth factors are added.  
     
     
         54 . Culture medium according to  claim 48 , characterized in that autologous growth factors and adhesion molecules are produced from platelets.  
     
     
         55 . Culture medium according to  claim 48 , characterized in that autologous growth factors and adhesion molecules are produced from leucocytes.  
     
     
         56 . Culture medium according to  claim 48 , characterized in that autologous growth factors and adhesion molecules are produced from platelets leucocytes.  
     
     
         57 . Culture medium according to  claim 48 , characterized in that autologous growth factors and adhesion molecules are processed from autologous clotted whole blood by centrifugation technique.  
     
     
         58 . Culture medium according to  claim 57 , characterized in that the autologous whole blood is stored for at least 1 hour at 37° C.  
     
     
         59 . Culture medium according to  claim 57 , characterized in that the autologous whole blood is stored for at least for 6 hours at 4° C.  
     
     
         60 . Culture medium according to  claim 54  and  57 , characterized in that the autologous growth factors and adhesion molecules are gained from enriched (concentrated) platelets.  
     
     
         61 . Culture medium according to  claim 53 , characterized in that the recombinant growth factor is bFGF, VEGF, EGF, TGF, Scatter-factor, PDGF or the combination of these growth factors.  
     
     
         62 . Culture medium according to any of  claims 48  to  61 , further comprising glycosaminoglycane.  
     
     
         63 . Culture medium according to  claim 62 , characterized in that the glycosaminoglycane is heparine, heparinsulfae, chondroitin, chondroitinsulfate, dermatin or dermatinsulfate.  
     
     
         64 . Culture medium according to any of  claims 48  to  63 , further comprising transferrin.  
     
     
         65 . Culture medium according to any of  claims 48  to  63 , further comprising hydrocortisone.  
     
     
         66 . Culture medium according to any of  claims 48  to  63 , further comprising insuline.  
     
     
         67 . Culture medium according to any of  claims 48  to  63 , further comprising albumine.  
     
     
         68 . Culture medium according to any of  claims 48  to  63 , characterized in that the culture medium is used for the culture of vascular cells, in particular endothelial cells, pericytes, pericyte-like-cells and smooth muscle cells.  
     
     
         69 . Culture medium according to any of  claims 47  to  63 , characterized in that the culture medium is used for the culture of non-vascular cells, in particular from hepatocytes.  
     
     
         70 . Culture medium according to any of  claims 48  to  69 , characterized in that the culture medium is used for the precoating of vascular prostheses, cardiac valves and bypasses in tissue engineering.  
     
     
         71 . Culture medium according to any of  claims 48  to  70 , characterized in that the culture medium is used within the framework of tissue engineering.  
     
     
         72 . Culture medium according to any of  claims 48  to  71 , characterized in that the culture medium is used as a preservation solution in tissue banking.  
     
     
         73 . Culture medium according to any one of  claims 48  to  72 , characterized in that, that autologous growth factors are gained through mechanical destruction of body-own tissues.